A culture system for the live analysis of successive developmental processes and the morphological control of mammalian vertebral cartilage
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The mesoderm-derived segmental somite differentiates into dermomyotome and sclerotome, the latter of which undergoes vertebrogenesis to spinal cartilage and ultimately to vertebral bones. However, analysis and manipulation of the developing mammalian vertebrae in the same embryo has been infeasible because of their placental-dependent embryogenesis. Here, we report a novel culture system of the mouse embryonic tailbud, by which the developmental processes of mammalian vertebral cartilage are traceable and manipulatable in the same sample. The anaplastic segmental somites/sclerotomes in the tailbud of 13 gestational day (g.d.) embryo that are structurally continuous to the vertebral column underwent progressive vertebrogenesis when (1) the ectoderm-derived nascent epidermis was microsurgically removed prior to cultivation, and (2) the sample was incubated at the air-medium interface. After cultivation for 5 days, the size and shape of the instructed vertebral cartilage showed features comparable to well-differentiated body vertebra along with the expression of the cartilage marker collagen type II, suggesting that aggressive differentiation of the sclerotomal cell lineage was achieved. In the presence of recombinant bone morphogenic protein (BMP) and Noggin, or adenoviral particles for extracellular epimorphin, dramatic alteration of the vertebral morphology ensued in the explants. Thus, this model system provides an approach to study the detailed molecular mechanisms of mammalian vertebrogenesis and enables pretreatment strategies of precartilagious fragments for improving the efficacy of subsequent transplantation.
KeywordsVertebrogenesis Live analysis Cartilage Epimorphin Organ culture Morphogenesis
We are grateful to Dr. Bascom for the critical reading of the manuscript and to all members of the Hirai laboratory for helpful discussions. Part of this work was supported by Hyogo COE Program Promotion Project.
- Farr J 2nd (2008) Autologous chondrocyte implantation and anteromedialization in the treatment of patellofemoral chondrosis. Orthop Clin North Am 39:329–335, viGoogle Scholar
- Balling R, Lau CF, Dietrich S, Wallin J, Gruss P (1992) Development of the skeletal system. Ciba Found Symp 165:132–140; discussion 140-133Google Scholar
- Fuchs JR, Terada S, Ochoa ER, Vacanti JP, Fauza DO (2002) Fetal tissue engineering: in utero tracheal augmentation in an ovine model. J Pediatr Surg 37:1000–1006; discussion 1000-1006Google Scholar
- Hirai Y, Nose A, Kobayashi S, Takeichi M (1989a) Expression and role of E- and P-cadherin adhesion molecules in embryonic histogenesis. I. Lung epithelial morphogenesis. Development 105:263–270Google Scholar
- Hirai Y, Nose A, Kobayashi S, Takeichi M (1989b) Expression and role of E- and P-cadherin adhesion molecules in embryonic histogenesis. II. Skin morphogenesis. Development 105:271–277Google Scholar
- Lippert H (1966) Anatomy of the spine from the viewpoints of development and function. Med Klin 61:41–46Google Scholar
- Marcelle C, Stark MR, Bronner-Fraser M (1997) Coordinate actions of BMPs, Wnts, Shh and noggin mediate patterning of the dorsal somite. Development 124:3955–3963Google Scholar
- Monsoro-Burq AH, Duprez D, Watanabe Y, Bontoux M, Vincent C, Brickell P, Le Douarin N (1996) The role of bone morphogenetic proteins in vertebral development. Development 122:3607–3616Google Scholar
- Shinohara H (1999a) The mouse vertebrae: changes in the morphology of mouse vertebrae exhibit specific patterns over limited numbers of vertebral levels. Okajimas Folia Anat Jpn 76:17–31Google Scholar
- Shinohara H (1999b) The musculature of the mouse tail is characterized by metameric arrangements of bicipital muscles. Okajimas Folia Anat Jpn 76:157–169Google Scholar